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Team:NTNU Trondheim/Characterization
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==Characterization Results== | ==Characterization Results== | ||
| - | To be able to use the rrnB P1 promoter, most of the BioBrick BBa_K112118 (lacking the first 11 bp) was amplified with PCR. The primers were designed to give the | + | To be able to use the rrnB P1 promoter, most of the BioBrick BBa_K112118 (lacking the first 11 bp) was amplified with PCR. The primers were designed to give the [Openwetware prefix and suffix], which contain E, X and S, P restriction sites. |
To test the downregulating-effect of ppGpp on rrnB P1, which is the basis of our project, we decided to make a construct containing ppGpp Synthase (RelA) inducible by the pBAD/AraC promoter. The construct would also contain beta-galactosidase (lacZ) which would be expressed by the rrnB P1 promoter. | To test the downregulating-effect of ppGpp on rrnB P1, which is the basis of our project, we decided to make a construct containing ppGpp Synthase (RelA) inducible by the pBAD/AraC promoter. The construct would also contain beta-galactosidase (lacZ) which would be expressed by the rrnB P1 promoter. | ||
Thus, induction of the pBAD/AraC promoter with arabinose should give lower lacZ production, as relA is overproduced giving high ppGpp concentration. | Thus, induction of the pBAD/AraC promoter with arabinose should give lower lacZ production, as relA is overproduced giving high ppGpp concentration. | ||
Revision as of 11:58, 22 August 2011
Characterization Results
To be able to use the rrnB P1 promoter, most of the BioBrick BBa_K112118 (lacking the first 11 bp) was amplified with PCR. The primers were designed to give the [Openwetware prefix and suffix], which contain E, X and S, P restriction sites.
To test the downregulating-effect of ppGpp on rrnB P1, which is the basis of our project, we decided to make a construct containing ppGpp Synthase (RelA) inducible by the pBAD/AraC promoter. The construct would also contain beta-galactosidase (lacZ) which would be expressed by the rrnB P1 promoter. Thus, induction of the pBAD/AraC promoter with arabinose should give lower lacZ production, as relA is overproduced giving high ppGpp concentration.
